title: Hemoglobin Warsaw (Phe beta 42(CD1)----Val), an unstable variant with decreased oxygen affinity. Characterization of its synthesis, functional properties, and structure.
abstract: In Hb Warsaw Val replaces the Phe normally present at the heme contact position beta 42 (CD1). This variant is unstable, and it readily undergoes methemoglobin formation. In DEAE-cellulose chromatography, the variant hemoglobin co-eluted with Hb A; a partially heme-depleted fraction of the variant, representing 5-6% of the total hemoglobin, eluted separately and in pure form. The heme replete form of Hb Warsaw exhibited decreased oxygen affinity with a normal Bohr effect and normal cooperativity and interaction with 2,3-diphosphoglycerate (DPG). The heme-depleted Hb Warsaw had a higher oxygen affinity than that of Hb A, decreased cooperativity and 2,3-DPG interaction, and a very low alkaline Bohr effect. Gel filtration of the heme-depleted form showed it to exist entirely as alpha beta dimers. Globin chain synthesis by Hb Warsaw-containing reticulocytes followed a balanced alpha/beta ratio. In short-term synthesis experiments, a major portion of incorporated radiolabeled L-leucine was recovered from the dimeric, heme-depleted Hb Warsaw fraction, suggesting that subunit association precedes the incorporation of heme into the beta subunits in the post-synthetic assembly of this hemoglobin. Structural analysis of deoxyhemoglobin containing roughly equal proportions of normal and variant beta chains showed that the replacement leaves a cavity next to the heme that is large enough to hold a water molecule, which may account for the instability of Hb Warsaw. The heme and the pyrrol nearest to ValCD1 tilt into the cavity. The resulting increase in the tilt of the proximal histidine relative to the heme plane, coupled with a possible stretching of the Fe-N epsilon bond may account for the low oxygen affinity.
authors.affiliation: Department of Pediatrics, University of Illinois College of Medicine, Chicago 60612.
authors.forename: G R
authors.initials: GR
authors.lastname: Honig
authors.forename: L N
authors.initials: LN
authors.lastname: Vida
authors.forename: B B
authors.initials: BB
authors.lastname: Rosenblum
authors.forename: M F
authors.initials: MF
authors.lastname: Perutz
authors.forename: G
authors.initials: G
authors.lastname: Fermi
mesh_terms: D019794:2,3-Diphosphoglycerate; D002848:Chromatography, DEAE-Cellulose; D002850:Chromatography, Gel; D004163:Diphosphoglyceric Acids / Q000378:metabolism; D005914:Globins / Q000096:biosynthesis; D006455:Hemoglobins, Abnormal / Q000302:isolation & purification / Q000502:physiology*; D006801:Humans; D046911:Macromolecular Substances; D008706:Methemoglobin / Q000378:metabolism; D015394:Molecular Structure; D010100:Oxygen / Q000097:blood*; D010649:Phenylalanine; D013329:Structure-Activity Relationship; D014633:Valine
chemical_list: D004163:Diphosphoglyceric Acids; D006455:Hemoglobins, Abnormal; D046911:Macromolecular Substances; C050261:hemoglobin Sendagi; D019794:2,3-Diphosphoglycerate; D010649:Phenylalanine; D005914:Globins; D008706:Methemoglobin; D014633:Valine; D010100:Oxygen